脐带血单个核细胞体外细胞因子诱导培养前后淋巴细胞亚群的变化及意义  

The significance and change of lymphocyte subsets of the mononuclear cells from umbilical cord blood before and after induction culture by cytokines in vitro

作　　者：董晋豫 刘建雷 曾赤佳 余俊健 邬素珍 刘劼 DONG Jin-yu;LIU Jian-lei;ZENG Chi-jia(Department of Clinical Laboratory,Center Hospital of Chancheng District,Guangdong Foshan 528000)

机构地区：[1]广东省佛山市禅城区中心医院检验科,广东佛山528000 [2]广东省佛山市禅城区中心医院中医妇科,广东佛山528000

出　　处：《医学检验与临床》2020年第11期5-9,共5页Medical Laboratory Science and Clinics

基　　金：广东省佛山市医学类科技攻关项目(2017AB002391)。

摘　　要：目的:探讨脐带血单个核细胞经细胞因子诱导培养前后淋巴细胞亚群的变化。方法:分离脐带血单个核细胞,按照细胞因子诱导的杀伤细胞(cytokine induced killer cells,CIK)的培养方法,加入rhlFN-y、rhIL-2、抗CD3和CD28单克隆抗体进行体外诱导培养14天,然后采用流式细胞仪检测诱导前后的淋巴细胞亚群比例。结果:诱导培养14d后的CIK细胞中T细胞(CD3+)、杀伤性T细胞(CD3+CD8+)、CD3+CD56+细胞比例均比诱导前显著增加[(97.67±1.07)%VS(62.47±11.67)%,P<0.01],[(58.55±13.70)%VS(15.72±6.03)%,P<0.01],[(26.68±9.80)%VS(0.64±0.70)%,P<0.01],而辅助性T细胞(CD3+CD4+)、B细胞(CD3-CD19+)、NK细胞(CD3-CD16+56+)比例均比诱导前下降[(37.29±14.03)%VS(45.87±10.96)%,P>0.05],[(0.95±1.17)%VS(29.85±11.29)%,P<0.01],[(1.95±1.08)%VS(16.52±8.68)%,P<0.01]。CD3+CD8+T细胞中,中央型记忆T细胞(CD45RO+CD62L+)和效应型记忆T细胞(CD45RO+CD62L)、中期分化T细胞(CD27+CD28)、活化T细胞(CD38+HLA-DR+)比例均较诱导前增加[(47.48±20.52)%VS(4.32±3.48)%,P<0.01]、[(52.08±20.60)%VS(2.05±1.47)%,P<0.01],[(79.12±9.24)%VS(3.49±1.57)%,P<0.01],[(27.54±19.24)%VS(1.32±2.12)%,P<0.01],差异均有统计学意义。结论:rhIFN-Y、rhIL-2、抗CD3和CD28单克隆抗体诱导培养的脐带血CIK细胞中的主要细胞群是CD3+CD56+细胞、CD3+CD8+T细胞和CD3+CD4+T细胞,并且以中期分化的记忆T细胞为主。Objective:To investigate the change of lymphocyte subsets of the mononuclear cells from umbilical cord blood before and after induction culture by cytokines in vitro.Methods:The mononuclear cells isolated from umbilical cord blood were induced to the cultivation of CIK cells for 14 days by adding rhIFN-γ,rhIL-2,anti-CD3 and CD28 monoclonal antibodies.The change of lymphocyte subsets before and after induction was detected by flow cytometry.Results:The proportion of T cells(CD34),killer T cells(CD3+CD8+)and CD3+CD56+cellls in CIK induced for 14 days was increased significantly than that before induction[(97.67±1.07)%VS(62.47±11.67)%,P<0.01],[(58.55±13.70)%VS(15.72±6.03)%,P<0.01],[(26.68±9.80)%VS(0.64±0.70)%,P<0.01].The proportion of helper T cells(CD3+CD4+),B cells(CD3+CD19+),NK cells(CD3-CD16+56+)was significantly lower than that before induction[(37.29±14.03)%VS(45.87±10.96)%,P>0.05],[(0.95±1.17)%VS(29.85±11.29)%,P<0.01],[(1.95±1.08)%VS(16.52±8.68)%,P<0.01].The ratio of central memory T cells(CD45RO+CD62L+) and effector memory T cells(CD45RO+CD62L-),mid-differentiation T cells(CD27+CD28-)and activated T cells(CD38+HLA-DR+)in CD8+T cells was increased significantly compared to that before induction[(47.48±20.52)%VS(4.32±3.48)%,P<0.01],[(52.08±20.60)%VS(2.05±1.47)%,P<0.01],[(79.12±9.24)%VS(3.49±1.57)%,P<0.01],[(27.54±19.24)%VS(1.32±2.12)%,P<0.01],the difference was statistically significant.Conclusion:The main cells in CIK cells from cord blood induced by rhIFN-Y,rhIL-2,anti-CD3 and CD28 monoclonal antibodies are CD3+CD56+,CD3+CD8+and CD3+CD4+T cells,which are mainly memory T cells with medium-term differentiation.

关 键 词：脐带血 单个核细胞 细胞因子 淋巴细胞亚群 

分 类 号：R73[医药卫生—肿瘤]